马钱子碱经皮给药对乳腺癌骨转移抑制机制的探讨

目的:探讨马钱子碱经皮给药对乳腺癌骨转移的抑制机制,为乳腺癌的治疗提供新的途径.方法:培养人乳腺癌细胞株MDA-MB-231,取对数生长期细胞制成单细胞悬液.20只雌性Balb/c-nu/nu裸鼠麻醉后,无菌条件下,局部骨内注射乳腺癌单细胞悬液0.2 mL,复制乳腺癌骨转移模型.20只荷瘤裸鼠随机分为马钱子碱高剂量组、中剂量组、低剂量组和模型组4组,分别给予20％(0.2 mg/g)、10％(0.1 mg/g)和5％(0.05 mg/g)浓度的马钱子碱膏(每日总剂量≤腹腔给药的半数致死量)和凡士林经皮给药,涂抹裸鼠的整个腹部和造模的部位,1 g/次,4次/d(每次间隔3 h),连续15 d.影像学观察骨转移情况;RT-PCR检测并比较各组肿瘤组织中,骨转移相关因子VEGF、COX-2和PTHrP的表达量.结果:胫骨的X射线表现,马钱子碱高剂量组和中剂量组的骨质破坏不明显,低剂量组和模型组的骨质破坏明显,甚至出现病理性骨折.RT-PCR结果显示,钱子碱高剂量组VEGF、COX-2和PTHrP mRNA的表达分别为25.30±0.90、20.63±0.73和25.56±0.54,马钱子碱中剂量组分别为24.10±0.73、20.49±0.76和25.74±0.48,马钱子碱低剂量组分别为25.27±0.75、21.02±0.65和25.65±0.44,模型组分别为24.09±0.74、21.14±0.66和25.75±0.39.与模型组比较,马钱子碱各剂量组VEGF、COX-2和PTHrP mRNA的表达量随着马钱子碱剂量的增加逐渐下降,差异均有统计学意义,P＜0.05.结论:马钱子碱经皮给药能抑制裸鼠乳腺癌骨转移瘤的生长,减轻骨损伤,其机制可能是通过调控VEGF、COX-2和PTHrP的表达而抑制乳腺癌骨转移瘤的生长.     

乳腺癌MRMT-1细胞局部骨转移大鼠模型的研究

目的 研究乳腺癌MRMT-1细胞局部骨转移大鼠模型在痛觉反应、影像学、病理学和分子生物学等方面的表现,为进一步的发病机制和药理学研究提供资料.方法 将32只雌性SD大鼠用随机数字表法分为假手术组和模型组各16只,在模型组大鼠胫骨内注射乳腺癌MRMT-1细胞,引起局部骨转移.造模后19d测定痛觉反应,21d处死大鼠取材,测定肿瘤体积,用X线对骨质损伤进行评分并测定骨密度（BMD）,HE染色观察病理形态改变,抗酒石酸酸性磷酸酶（TRAP）法染色观察破骨细胞,免疫组织化学法检测核因子κB受体活化因子配体（RANKL）和骨保护素（OPG）水平,并计算比值;实时荧光定量反转录聚合酶链反应（RT-PCR）法检测甲状旁腺激素相关蛋白（PTHrP）水平.结果 与假手术组比较,模型组存在痛觉异常状态（P＜0.01）;胫骨在X线下损伤评分较高（P＜0.01）.模型组和假手术组大鼠左侧胫骨的BMD分别为（0.11 ±0.01）g/cm2和（0.13±0.02）g/cm2（P＜ 0.05）.模型组胫骨标本上肿瘤生长明显,体积为（1082.73±679.44）mm3,而假手术组无肿瘤生长（P＜0.01）,病理可见混合性骨转移,但以溶骨性病变为主;伴有破骨细胞数量明显增多,达到（40.84±25.59）个/高倍视野,而假手术组仅为（1.88±2.92）个/高倍视野（P＜0.01）.转移灶局部RANKL水平无明显变化,OPG水平下降（P＜0.05）,OPG/RANKL比值和PTHrP下降（P＜0.05）.结论 乳腺癌MRMT-1细胞局部骨转移大鼠模型存在癌痛、骨质破坏,甚至病理性骨折等表现.其发病机制可能是MRMT-1细胞在浸润生长过程中打破了OPG-RANKL-RANK系统的平衡,从而激活破骨细胞,引起骨吸收作用亢进,引起各种表现.     

生酮饮食对小鼠EMT-6乳腺癌生长影响的实验研究

目的 研究生酮饮食(ketogenic diet,KD)对小鼠EMT-6乳腺癌生长的影响及其机制.方法 取40只BALB/c雌性小鼠,建立小鼠EMT-6乳腺癌模型后,随机平均分配到4个饮食组,每组10只,分别给予标准饮食(standard diet,SD),20％糖生酮饮食(20％糖KD),10％糖生酮饮食(10％糖KD)及无糖生酮饮食(NCKD)饲养.观察小鼠肿瘤生长情况、生存时间及血糖和血酮的变化.采用酶联免疫吸附分析法(ELISA)检测血胰岛素和血胰岛素样生长因子-1(insulin-like growth factors-1,IGF-1)的变化.当小鼠肿瘤体积超过1 000 mm3时即为观察终点,处死小鼠.从接种到观察终点之间的天数记为小鼠生存时间.结果 接种后第16天,各KD组肿瘤体积显著小于SD组,SD组肿瘤平均体积为(319.59±36.03) mm3,20％糖KD组为(205.11±42.68) mm3,10％糖KD组为(217.03±35.80) mm3,NCKD组为(248.36±44.01) mm3,差异有统计学意义,F=176.123,P＜0.001;各KD组与SD组之间两两比较差异均有统计学意义,P值均＜0.001.接种后第7天各KD组血糖水平较SD组降低,SD组为(6.16±0.33) mmol/L,20％糖KD组为(2.93±0.20) mmol/L,10％糖KD组为(3.02±0.26) mmol/L,NCKD组为(2.89±0.17) mmol/L,F=421.529,P＜0.001;各KD组与SD组之间两两比较差异均有统计学意义,P＜0.001.接种后第7天各KD组血酮水平较SD组升高,SD组为(0.60±0.15) mmol/L,20％糖KD组为(2.54±0.25) mmol/L,10％糖KD组为(2.67±0.34) mmol/L,NCKD组为(2.65±0.16) mmol/L,F=183.395,P＜0.001;各KD组与SD组之间两两比较差异均有统计学意义,P＜0.001.SD组、20％糖KD组、10％糖KD组和NCKD组的中位生存时间分别为26、34、32和32 d,Log-rank检验显示,SD组与20％糖KD组差异有统计学意义,x2=14.044,P＜0.001;SD组与10％糖KD组差异有统计学意义,x2=10.199,P=0.001;SD组与NCKD组差异有统计学意义,x2=8.038,P=0.005.运用多因素Cox比例风险回归模型分析接种后第14天各KD组血糖、血酮及体质量增长量发现,血糖(P=0.026,RR=2.548,95％CI为1.062～3.064)及血酮(P=0.013,RR=0.272,95％CI为0.059～0.503)均对生存有影响.结论 生酮饮食可抑制小鼠EMT-6乳腺癌的生长,其机制与血糖及血酮的变化相关.     

乳腺癌多药耐药DEDD作用及机制研究

目的 死亡效应结构域DNA结合蛋白(death effector domain DNA-binging protein,DEDD)可抑制乳腺癌的生长和转移,但在乳腺癌多药耐药中的作用尚不明确.本研究将探讨DEDD在人乳腺癌多药耐药中的作用及机制.方法 通过转染DEDD-shRNA建立稳定敲低DEDD表达的MCF-7人乳腺癌细胞株,MTS实验检测多柔比星、紫杉醇对MCF-7 WT(Wild type,野生型)、MCF-7 control shRNA和MCF-7 DEDD-shRNA细胞活力的影响,Annexin-V和碘化丙啶(propidium iodide,PI)双染后流式细胞仪检测细胞凋亡.蛋白质印迹法检测乳腺癌耐药蛋白(breast cancer resistance protein,BCRP)在药物干预后的蛋白表达水平.结果 多柔比星干预后,半数抑制浓度IC50的多因素方差分析结果显示,不同处理(F=89.49,P＜0.001)、不同时间(F=50.81,P＜0.001)组间比较差异有统计学意义.多柔比星干预24或48 h后,MCF-7 DEDD-shRNA的半数抑制浓度IC50分别为(0.97±0.15)和(0.62±0.09) μmol/L,明显高于MCF-7 WT(0.46±0.05)和(0.26±0.03) μmol/L和MCF-7 control shRNA细胞(0.39±0.05)和(0.25±0.03)μmol/L,P＜0.001.紫杉醇干预后,半数抑制浓度IC50的多因素方差分析结果显示,不同处理(F=15.94,P＜0.001)、不同时间组间(F=129.70,P＜0.001)比较差异有统计学意义.紫杉醇干预24或48 h后,MCF-7 DEDD-shRNA的半数抑制浓度IC50为(16.74±2.26)和(9.88±1.47) μmol/L,明显高于MCF-7 WT(13.39±1.44)和(7.69±0.92) μmol/L(P=0.001)和MCF-7 control shRNA细胞(12.58±1.15)和(7.17±1.12) μmol/L(P＜0.001).多柔比星干预后24 h后流式细胞凋亡结果显示,不同处理(F=131.46,P＜0.001)、不同浓度组间(F=160.95,P＜0.001)比较差异有统计学意义.正常培养状态下,MCF-7 control shRNA细胞和MCF-7 DEDD-shRNA细胞的凋亡率分别为(14.32±1.47)％和(11.58±1.63)％,而给予0.5μmol/L和1 μmol/L多柔比星作用24 h后,M[CF-7DEDD-shRNA细胞的凋亡率分别为(21.62±1.97)％和(24.39±2.36)％,明显低于MCF-7 control shRNA细的(36.26±1.87)％和(38.23±1.46)％,P＜0.001.同样,紫杉醇干预后24 h,不同处理(F=124.81,P＜0.001)、不同浓度组问(F=172.56,P＜0.001)细胞凋亡率比较差异有统计学意义.给予9.37和18.74 μmol/L紫杉醇作用24 h后,MCF-7DEDD-shRNA细胞的凋亡率分别为(30.26±1.63)％和(32.18±2.16)％,明显低于MCF-7 control shRNA组的(53.84±2.17)％和(58.27±2.16)％,P＜0.001.多柔比星作用后不同时间BCRP蛋白质印迹检测统计分析结果显示,不同处理(F=14.67,P＜0.001)、不同时间(F=6.39,P=0.006)组间BCRP蛋白表达比较差异有统计学意义.给予0.5 μmol/L多柔比星作用24 h后,3组细胞的BCRP相对表达水平分别为0.87±0.04、1.06±0.02和5.25±0.18.MCF-7 DEDD-shRNA细胞组BCRP蛋白的表达水平显著高于MCF-7 WT和MCF-7 control shRNA细胞组,P＜0.001.多柔比星作用48 h后,3组细胞的BCRP相对表达水平分别为1.06±0.01、0.97±0.04和2.98±0.13.MCF-7 DEDD-shRNA细胞组BCRP蛋白的表达水平出现回落,但仍显著高于MCF-7 WT和MCF-7 control shRNA细胞组,P＜0.001.同样,紫杉醇作用后不同时间BCRP蛋白质印迹检测统计分析结果显示,不同处理(F=7.26,P=0.004)、不同时间(F=8.32,P=0.002)组间BCRP蛋白表达比较差异有统计学意义.给予9.37 μmol/L紫杉醇作用24 h后,3组细胞的BCRP相对表达水平分别为0.81±0.05、2.25±0.10和1.78±0.14.其中MCF-7 control shRNA组细胞BCRP表达水平高于MCF-7 WT (P＜0.001)和MCF-7 DEDD-shRNA (P=0.002)组.而紫杉醇作用48 h后,3组细胞的BCRP相对表达水平分别为0.73±0.04、1.73±0.05和3.12±0.12,MCF-7 DEDD-shRNA细胞组BCRP蛋白的表达水平继续增高,且显著高于MCF-7 WT和MCF-7 control shRNA细胞组,P＜0.001.结论 敲低DEDD表达可增强乳腺癌细胞的抗肿瘤药耐药性,BCRP的表达增加可能是低表达DEDD乳腺癌细胞多药耐药的机制之一.     

抑制肿瘤细胞骨保护素表达对乳腺癌细胞致骨转移能力的影响

目的观察抑制肿瘤细胞骨保护素（0steoprotegerin,OPG）表达对乳腺癌MDA～MB-231细胞株致骨转移的影响。方法32只4～6周龄雌性裸鼠被随机分为A、B、C、D4组,每组8只,A、C组的每只裸鼠按分组分别以5×10。个MDA—MB-231细胞注射入左心室（A组）或左胫骨骨髓腔（C组）。B、D组的每只裸鼠按分组分别以5×10。个MDA—MB-231i细胞（OPG表达抑制）注射人左心室（B组）或左胫骨骨髓腔（D组）,42d后行病理检查,比较A、B组骨转移发生率和骨转移灶数量,以及C、D组的骨肿瘤体积。定量资料的比较采用t检验或秩和检验,定性资料比较采用Fisher确切概率检验。结果A组有6只发生骨转移,全组共检出不连续性骨转移灶23处;B组有3只发生骨转移,全组共检出不连续性骨转移灶8处。A组的骨转移发生率和转移灶数量虽高于B组,但两者差异并无统计学意义（P〉0．05）。C组肿瘤平均体积为（66．29±41．01）mm3,D组肿瘤平均体积为（23．70±16．14）mm3,C组骨肿瘤体积大于D组,两组间差异有统计学意义（P=0．02）。结论乳腺癌细胞致骨转移能力与其OPG表达水平密切相关,抑制OPG表达可以降低乳腺癌细胞致骨转移的能力。     

淋巴管密度在子宫内膜癌淋巴结转移中的作用

[目的] 检测月经周期正常子宫内膜层和肌层以及子宫内膜癌组织中淋巴管密度(LVD),分析其与子宫内膜癌淋巴结转移的相关性.[方法] 收集正常子宫全层标本30例(15例增殖期,15例分泌期),子宫内膜癌标本40例(20例Ⅰa期,20例Ⅲc期),应用 D2-40 抗体进行免疫组化染色,显微镜下计数 LVD.[结果] 在正常月经周期的子宫内膜层和肌层均有D2-40 阳性染色的淋巴管,淋巴管与螺旋小动脉伴行.正常肌层 LVD 显著高于内膜层(10.70±1.16/mm2 vs 4.98±0.84/mm2; P＜0.01);癌灶内部 LVD(Ⅰa期3.49±0.94/mm2,Ⅲc期2.43±1.08/mm2)较正常内膜层和肌层均有所降低,差异显著(P＜0.01).而癌灶周边 LVD 升高,高于正常内膜但是低于正常肌层(P＜0.05).有淋巴结转移组癌灶周边 LVD (8.63±2.88/mm2)较未转移组显著增高(7.09±1.22/mm2,P=0.012).[结论] 癌灶周边淋巴管与子宫内膜癌的淋巴结转移发生有关.     

乳腺癌术后患者三氧治疗对血清TK1水平变化的影响

目的:探讨切除术后乳腺癌患者三氧治疗前后血清胸苷激酶 1(thymidine kinase-1,TK1)水平的变化及其与肿瘤分期的关系.方法:应用酶免疫点印记化学发光法检测104例切除术后乳腺癌患者三氧疗法前后血清TK1水平.比较不同临床分期乳腺癌患者接受三氧疗法前后 TK1、CA15-3及CEA水平的差异,分析TK1与雌激素受体(ER)和孕激素受体(PR)的关系.结果:三氧治疗前乳腺癌患者TK1平均值为(1.52±1.53) pmol/L,与健康体检组的(0.54±0.42) pmol/L相比,差异有统计学意义(P=0.000);三氧治疗后TK1平均值为(0.73±0.82) pmol/L,与治疗前相比,差异有统计学意义(P=0.001).根据临床分期,Ⅰ-Ⅲ期患者治疗前TK1平均值为(1.05±1.17) pmol/L,治疗后平均值为(0.38±0.44) pmol/L,差异有统计学意义(P=0.002);Ⅳ期患者治疗前TK1平均值为(2.53±1.73) pmol/L,治疗后平均值为(1.47±0.94) pmol/L,差异无统计学意义(P=0.080).CA15-3及CEA在三氧治疗前后比较,差异均无统计学意义,但Ⅰ-Ⅲ期患者CA15-3及CEA与TK1变化趋势有一致性.TK1与ER、PR有关联(r=0.455,P=0.000).结论:三氧治疗能有效降低切除术后乳腺癌患者的TK1水平,抑制肿瘤细胞增殖,控制疾病进展,且对Ⅰ-Ⅲ 期患者更有意义.     

凉血疏肝方治疗雌激素受体阳性乳腺癌术后患者的疗效分析

目的探讨凉血疏肝方对雌激素受体阳性乳腺癌术后患者的影响。方法选取80例经病理检查证实为雌激素受体阳性乳腺癌术后的患者,按照治疗方案将患者分成两组(每组各40例),对照组予以枸橼酸他莫昔芬片治疗,观察组在治疗组的基础上加用凉血疏肝方治疗,观察两组患者治疗前后雌二醇水平和子宫内膜厚度,治疗后两组患者血细胞分析、肝功能、肾功能有无异常,治疗3个月后是否出现复发转移。结果治疗前两组患者雌二醇水平比较无差异[(243.78±54.50)pmol/L vs(252.16±63.35)pmol/L](P>0.05);治疗后观察组雌二醇水平明显高于对照组[(128.44±56.34)pmol/L vs(145.97±59.39)pmol/L](P<0.05);治疗前两组患者子宫内膜厚度比较无差异[(5.22±1.65)mm vs(5.86±1.76)mm](P>0.05),治疗后对照组子宫内膜厚度明显增厚(P<0.05),观察组子宫内膜厚度变薄(P<0.05),两组患者比较有差异[(4.27±1.43)mm vs(7.74±1.48)mm](P<0.05);两组均未出现严重的骨髓抑制、肝功能异常及肾功能异常,并且均未出现乳腺癌转移复发的情况。两组患者不良反应比较差异有统计学意义(χ~2=6.135,P=0.013)。结论凉血疏肝方能有效减轻枸橼酸他莫昔芬治疗引起的副作用,并且无明显毒副作用,值得在临床上推广使用。     

乳腺癌骨扫描诊断骨转移特点分析

目的 :了解乳腺癌骨转移的特点。方法 :通过对 93例乳腺癌骨转移患者的骨扫描结果进行回顾性分析 ,了解乳腺癌骨转移的特点。结果 :乳腺癌患者骨转移表现以异常放射性浓集灶为主 ,部分患者也可表现为放射性稀疏缺损区。骨转移病灶以脊柱及肋骨为最多 ,占 80 % ;颅骨、胸骨、骨盆及四肢骨约占 2 0 %。浸润性导管癌最多 ,占 3 5 5 % ;腺癌其次 ,占 2 5 8% ;髓样癌占 12 9% ;黏液癌仅占 4 3 %。结论 :乳腺癌患者骨转移表现以异常放射性浓集灶为主 ,部分患者也可表现为放射性稀疏缺损区 ;浸润性导管癌和腺癌骨转移率高于髓样癌和黏液癌     

miR-342与乳腺癌ERα表达及他莫昔芬敏感性关系探讨

目的:探讨miR-342与乳腺癌临床病理的关系及是否参与调节ERα并影响他莫昔芬(TAM)的敏感性。方法:RT-PCR检测48例癌组织miR-342和ERαmRNA水平及其中24例癌旁组织miR-342的表达;免疫组化评价癌组织ER、PR、HER-2和VEGF的状态;RT-PCR检测乳腺癌细胞株MCF-7、SKBR-3及MB-231的miR-342、ERαmRNA水平;检测MCF-7细胞瞬时转染hsa-miR342(分mimic、inhibitor及各自阴性对照共4组)48h后miR-342和ERαmRNA的变化;1×10-8 mol/L 17-β雌二醇(E2)单独或联合2×10-5 mol/L TAM处理MCF-7细胞72h,CCK-8法测不同转染组细胞增殖;各转染组细胞1.5×10-5 mol/L TAM处理48h,流式细胞术分析细胞凋亡率。结果:miR-342及ERαmR-NA在ERα阳性乳腺癌组织及细胞中均明显升高,P<0.01;miR-342和ERαmRNA之间存在正相关,P=0.003。miR-342在HER-2阴性(P=0.001)及VEGF阴性(P=0.031)乳腺癌组织中上调;miR-342与PR、淋巴转移、病理分级等因素无明显关系,P>0.05;癌与癌旁miR-342表达差异无统计学意义,P=0.065。MCF-7细胞mimic组ERαmRNA表达为1.80±0.14,高于对照组的1.0±0.0,P=0.001;inhibitor组为0.747±0.087,较对照组低,P=0.037。TAM作用72h,mimic组细胞增殖率为(45.9±1.3)%,与对照组的(55.0±1.5)%相比明显抑制,P=0.001;inhibitor组增殖率为(72.9±1.9)%,较对照组升高,P=0.000。流式细胞术分析TAM处理后的细胞凋亡率,结果显示,mimic组凋亡率为(9.54±1.14)%,较对照组的(4.50±0.46)%增加,P=0.002;inhibitor组凋亡率为(3.06±0.42)%,较对照组的(4.95±0.59)%降低,P=0.011。结论:miR-342的表达可以一定程度预测ERα的表达水平,并作为分子标志预测乳腺癌细胞对TAM的敏感性;miR-342有望成为潜在靶点来参与乳腺癌内分泌治疗。     

Response to intravenous bisphosphonate therapy in hypercalcaemic patients with and without bone metastases: the role of parathyroid hormone-related protein.

Plasma parathyroid hormone related-protein (PTHrP) may inhibit the calcium-lowering effect of bisphosphonate therapy. In this prospective study we examined the relationship between plasma PTHrP levels, renal tubular markers of calcium reabsorption, and the effectiveness of intravenous bisphosphonate therapy (IVBPT) in lowering serum calcium in patients with hypercalcaemia of malignancy (HM), with and without bone metastases. Thirty-five symptomatic hypercalcaemic patients (17 without and 18 with bone metastases) were treated with IVBPT (pamidronate 30-60 mg or BM21.0955 2-6 mg). Normocalcaemia was achieved in 24/35 (71%) patients with a mean fall in serum calcium of 0.85 mmol l-1 (range 0.11-1.93, P < 0.001). In the 35 patients studied, serum calcium levels reached a nadir between days 3 and 7, and this was accompanied by a small but significant reduction in plasma PTHrP levels (median reduction 0.77 pmol l-1, P = 0.007). Patients who responded to bisphosphonate therapy by becoming normocalcaemic had significantly lower basal plasma PTHrP levels, mean 4.06 vs 8.2 pmol l-1 (P < 0.04). A significant reduction in urinary calcium excretion was seen (mean 106 mumol l-1, P < 0.02) in patients with bone metastases, and urinary cAMP (mean 170 mmol l-1, P < 0.01) fell in all patients. Patients without demonstrable bone metastases had significantly higher plasma PTHrP levels (P < 0.002), required more doses of IVBPT, and had a poorer reduction in serum calcium compared with patients with bone metastases, only one of whom required more than one dose. We conclude that circulating PTHrP has an important role in increasing renal tubular reabsorption of calcium in HM, thus reducing the effectiveness of bisphosphonate therapy, particularly in patients with humoral HM and no bone metastases.     

TGF-beta promotion of Gli2-induced expression of parathyroid hormone-related protein, an important osteolytic factor in bone metastasis, is independent of canonical Hedgehog signaling

Breast cancer frequently metastasizes to bone, in which tumor cells receive signals from the bone marrow microenvironment. One relevant factor is TGF-β, which upregulates expression of the Hedgehog (Hh) signaling molecule, Gli2, which in turn increases secretion of important osteolytic factors such as parathyroid hormone-related protein (PTHrP). PTHrP inhibition can prevent tumor-induced bone destruction, whereas Gli2 overexpression in tumor cells can promote osteolysis. In this study, we tested the hypothesis that Hh inhibition in bone metastatic breast cancer would decrease PTHrP expression and therefore osteolytic bone destruction. However, when mice engrafted with human MDA-MB-231 breast cancer cells were treated with the Hh receptor antagonist cyclopamine, we observed no effect on tumor burden or bone destruction. In vitro analyses revealed that osteolytic tumor cells lack expression of the Hh receptor, Smoothened, suggesting an Hh-independent mechanism of Gli2 regulation. Blocking Gli signaling in metastatic breast cancer cells with a Gli2-repressor gene (Gli2-rep) reduced endogenous and TGF-β-stimulated PTHrP mRNA expression, but did not alter tumor cell proliferation. Furthermore, mice inoculated with Gli2-Rep-expressing cells exhibited a decrease in osteolysis, suggesting that Gli2 inhibition may block TGF-β propagation of a vicious osteolytic cycle in this MDA-MB-231 model of bone metastasis. Accordingly, in the absence of TGF-β signaling, Gli2 expression was downregulated in cells, whereas enforced overexpression of Gli2 restored PTHrP activity. Taken together, our findings suggest that Gli2 is required for TGF-β to stimulate PTHrP expression and that blocking Hh-independent Gli2 activity will inhibit tumor-induced bone destruction.     

Parathyroid hormone-related protein localization in breast cancers predict improved prognosis

In a prospective study of 526 consecutive patients with operable breast cancer, the significance of positive parathyroid hormone-related protein (PTHrP) staining by immunohistology has been evaluated for a median of 10-year follow-up. Improved survival was observed for the 79% of tumors which stained positively for PTHrP [estimated univariate hazard ratio, 0.43; 95% confidence interval (95% CI), 0.30-0.62; P < 0.001]. Adjustments for N stage, progesterone receptor status, and log tumor size changed this estimate only slightly to 0.47 (95% CI, 0.63-0.69; P = 0.001). Patients with PTHrP-positive primary tumors were less likely to develop bone metastases (hazard ratio, 0.63; 95% CI, 0.41-0.98; P = 0.04). PTHrP status was associated with estrogen receptor (P = 0.01), progesterone receptor (P = 0.03), and menopausal status (P = 0.006) but was not significantly associated with tumor size, vascular invasion, tumor grade, or patient age. Of 19 patients requiring surgery for bone metastases, the primary cancers were PTHrP negative in seven, all but one of whom had PTHrP-positive bone metastases. All 12 patients with PTHrP-positive primary cancers also had positive bone metastases. We conclude that increased production of PTHrP by breast cancers confers on them a less invasive phenotype, an effect distinct from the bone resorption-stimulating action that favors bone metastasis. It is likely that the latter property is influenced by factors in the bone microenvironment.     

S-TK1在乳腺癌患者中检测的意义及其与预后的关系

目的研究S-TK1含量在乳腺癌、乳腺良性肿瘤患者及健康体检人群中的差别;探讨S-TK1与乳腺癌的关系,探讨乳腺癌患者S-TK1的含量与肿瘤分期、肿块大小、组织学分级、ER、PR及HER-2之间的关系;进而分析S-TK1含量与乳腺癌患者复发、转移的关系。方法采用化学发光点印迹法,分析乳腺癌、乳腺良性肿瘤患者及健康体检者S-TK1的含量。结果（1）乳腺癌患者治疗前（A组）、乳腺良性肿瘤患者（B组）及健康体检者（C组）S-TK1的含量分别为(17.98±10.20)pmol/L,(1.50±1.02)pmol/L,(0.88±0.45)pmol/L。A组S-TK1的含量明显高于B组和C组,差异有统计学意义（P均＜0.001）;B组S-TK1的含量高于C组,差异有统计学意义（P=0.017）。A组Ⅲ期乳腺癌患者S-TK1的含量（21.54±11.85 ）pmol/L高于Ⅱ期患者（14.05±6.19） pmol/L,差异有统计学意义（P=0.018）。（2）S-TK1的含量与组织学分级、HER-2及肿块大小均呈正相关（其相关系数分别为r＝0.596、0.324、0.397,对应的P值为P<0.001、=0.042、=0.011）;而ER与S-TK1的含量呈负相关（其r=－0.391,P=0.013）;PR与S-TK1的含量无相关性（P＝0.395,r=-0.138）。（3）复发转移组治疗前S-TK1含量(23.17±13.15) pmol/L明显高于无病生存组(14.31±5.50) pmol/L,差异有统计学意义（P=0.011）,S-TK1低值组和中值组无病生存高于S-TK1高值组,但差异无统计学意义（P=0.186）。结论S-TK1也许可以作为乳腺癌早期诊断的指标之一;S-TK1含量与乳腺癌患者的预后有关,治疗前高S-TK1含量可能更容易早期出现复发、转移。     

体素内不相干运动扩散加权成像对乳腺肿块样病变的诊断价值

目的 乳腺肿块样病变的诊断及鉴别对于临床治疗方案的制定及预后判断具有重要意义.本研究旨在探索体素内不相关运动(intravoxel incoherent motion,IVIM)扩散加权成像(diffusion weighted imaging,DWI)在乳腺肿块样病变中的诊断价值.方法 选取云南省肿瘤医院2015-07-01-2015-12-30乳腺肿块患者127例,共135个病灶,其中乳腺癌75例(75个病灶)、良性肿瘤27例(32个病灶)、囊肿10例(13个病灶)、炎性病变15例(15个病灶).所有患者行常规DWI及IVIM-DWI、动态增强(dynamic contrast-enhanced magnetic resonance imaging,DCE-MRI)检查.根据病理结果将病灶分为乳腺癌组、良性肿瘤组、囊肿组、炎性病变组,并以对侧正常腺体作为对照组.比较不同病变及正常腺体各组间ADC、D、D*和f值有无差异,评价上述各参数对乳腺肿块样良恶性病变的诊断效能.结果 在IVIM-DWI上,随b值增大,囊肿信号呈单指数线性衰减,正常腺体组织、炎性病变、良性肿瘤及乳腺癌信号呈双指数非线性衰减.单因素方差分析显示,各组ADC(F=119.35,P＜0.001)、D(F=58.31,P＜0.001)、f(F=50.961,P＜0.001)和D*值(F=2.732,P=0.032)比较差异有统计学意义;进一步两两比较,ADC值除对照组和良性肿瘤组外其余各组间差异均有统计学意义(P＜0.05),囊肿组ADC值最高,乳腺癌组最低;D值各组间两两比较差异均有统计学意义(P＜0.05);f值除炎性组和乳腺癌组、腺体组和良性肿瘤组外,其余各组间比较差异有统计学意义,P＜0.05;D*值仅囊肿组与其他4组比较差异有统计学意义,P＜0.05.各组D与ADC值比较,除囊肿组D值和ADC值差异无统计学意义外,余各组ADC值均高于D值,P＜0.001.D、ADC和f值诊断乳腺肿块样病变良恶性实验得到的曲线下面积(area under roc curve,AUC)分别为0.930、0.898和0.768,D值诊断效果最好,三者鉴别诊断乳腺肿块样良恶性病变的最佳临界值分别为0.89×10-3mm2/s、1.08×10-3 mm2/s和5.54％,相对应的诊断敏感性依次为95.7％、89.3％和79.2％,特异性依次为92.9％、87.5％和71.4％.结论 IVIM-DWI对于乳腺肿块样病变具有较高的诊断及鉴别诊断的价值,D值诊断效能较常规ADC值更好.     

Parathyroid hormone-related protein regulates the growth of orthotopic human lung tumors in athymic mice.

BACKGROUND: Parathyroid hormone-related protein (PTHrP) has growth regulatory effects for many malignant cells and may influence the progression of carcinomas of the breast, prostate, and lung. In the current study, the authors investigated the in vivo and in vitro effects of PTHrP neutralizing antibody and PTHrP treatment on the growth of BEN cells, a human lung squamous cell carcinoma line that expresses PTHrP and its receptor. METHODS: Orthotopic lung tumors were produced in 20 athymic mice with BEN-GFP cells (a clonal line that stably expresses green fluorescent protein [GFP]) by instilling suspensions of 3 x 10(6) cells per mouse into the lungs of anesthetized animals. The mice were divided into 2 groups receiving either subcutaneous mouse antihuman PTHrP antibodies or irrelevant mouse immunoglobulin (Ig) G (150 microg) twice weekly. RESULTS: After 30 days, 6 of 10 mice receiving anti-PTHrP antibodies had lung tumors visible on macroscopic inspection, but only 1 of the 10 mice treated with irrelevant IgG had a lung tumor that was of that size (P < 0.01). GFP fluorescence was significantly greater in lung homogenates of the PTHrP antibody-treated mice than in the mice treated with IgG (6006 +/- 411 vs. 2907 +/- 282 relative fluorescent units, respectively; P < 0.001). Although neutralizing antibodies stimulated BEN cell lung tumor growth, exogenous PTHrP 1-34 treatment (0.01-1 nM) inhibited the growth of cultured BEN cells by approximately 40%. CONCLUSIONS: Although PTHrP expression has been reported to be associated with more aggressive malignancies, the data from the current study suggest that PTHrP 1-34 was a paracrine growth inhibitor in BEN human lung carcinoma cells. The growth-related effects of PTHrP are complex, and can be both stimulatory and inhibitory.     

Osteoprotegerin overexpression by breast cancer cells enhances orthotopic and osseous tumor growth and contrasts with that delivered therapeutically

Osteoprotegerin (OPG) acts as a decoy receptor for receptor activator of NF-kappaB ligand (RANKL), which is a pivotal molecule required for osteoclast formation. In vitro OPG inhibits osteoclast formation and in vivo (administered as Fc-OPG) it reduces hypercalcemia and the establishment of osteolytic lesions in mouse models of tumor cell growth in bone. Osteolysis can be induced by parathyroid hormone-related protein (PTHrP) produced by breast cancer cells that results in an increased osteoblastic RANKL/OPG ratio. We examined the effect of local tumor production of OPG on the ability of breast cancer cells to establish and grow in bone and mammary fat pad. MCF-7 cells or MCF-7 cells overexpressing PTHrP were transfected with full-length OPG and inoculated into the proximal tibiae of athymic nude mice. Mice injected with cells overexpressing PTHrP and OPG showed enhanced tumor growth, increased osteolysis (2-fold compared with MCF-7 cells overexpressing PTHrP), and altered histology that was reflective of a less differentiated (more aggressive) phenotype compared with MCF-7 cells. In contrast, administration of recombinant Fc-OPG reduced tumor growth and limited osteolysis even in mice inoculated with OPG overexpressing cells. Similarly, OPG overexpression by breast cancer cells enhanced tumor growth following orthotopic inoculation. These results indicate that OPG overexpression by breast cancer cells increases tumor growth in vivo and that there are strikingly different responses between therapeutically administered Fc-OPG and full-length OPG produced by tumor cells.     

肺癌锥形束CT图像不同配准方式的误差分析

目的 研究肺癌锥形束CT(CBCT)图像配准的影响因素.方法 选取2007年间本科采用CBCT作为在线校位的肺癌放疗患者20例.每位患者在疗程中6～19次治疗采集了CBCT图像.根据肺内病变位置与椎体关系分为病变靠近椎体组(A组)和病变远离锥体组(B组),对比两组摆位误差差异.同时对该组病例分别选取从治疗初期、中期和后期CBCT图像,请4位医生分别采用骨配准和灰度配准,比较不同医生间和不同配准方式间差异.结果 A组和B组在头脚、左右、前后方向上的摆位误差分别为-1.31、1.24、-1.88 mm和0.10、1.37、-1.26 mm(t=0.07、0.05、-0.12,P=0.554、0.652、0.321).4位医生骨配准摆位误差头脚方向分别为-0.05、-0.01、0.05、-0.16(F=-0.01,P=0.887),左右方向分别为0.56、0.35、0.51、0.43 mm(F=-0.01,P=0.880),前后方向分别为-1.16、-1.20、-0.88、-1.03 mm(F=0.04,P=0.555);灰度配准摆位误差头脚方向分别为-0.32、-0.34、-0.39、-0.37 mm(F=-0.01,P=0.874),左右方向分别为0.34、0.54、-0.04、0.27 mm(F=-0.03,P=0.622),前后方向分别为-1.12、-1.15、-1.13、-1.04 mm(F=0.00,P=0.812).结论 采用相同配准框和图像质量下,肺癌患者肺内病变位置、不同配准方式和不同医生对图像引导放疗中CBCT图像配准无明显影响.

Abstract：

Objective To analyze the influencing factors of cone-beam CT (CBCT) imagine registration in lung cancer. Methods From Mar. 2007 to Dec. 2007, 20 patients with lung cancer were treated with IGRT. The imagines of CBCT were collected from 6 to 19 fractions during the patients' radiotherapy. To compare the difference of set-up errors between the two groups according to the distance from the lesion in lung to the centrum. At the same time, CBCT imagines from the first, middle and the last fraction of these patients' radiotherapy were registrated in bone and grey methods by four doctors. The difference of set-up errors between different doctors and registrated methods were compared. Results The mean values of set-up errors were ＜2 mm in the two groups without significant difference (x:-1.31mm vs 0. 10 mm (t=0. 07,P=0.554);y:1.24 mm vs 1.37 mm (t=0. 05,P=0. 652);z: - 1.88mm vs -1.26mm (t= -0. 12,P=0.321)). The mean values of set-up errors were ＜ 1.3 mm in four doctors and registrated methods without significant difference, for bone registration,x: -0. 05 mm, -0. 01 mm,0. 05 mm, -0.12 mm and -1.31 mm ( F=-0.01,P=0.887) ;y:0.56 mm,0.35 mm,0.51 mm and 0.43 mm (F= -0.01,P=0.880);z: -1.16 mm, -1.20 mm, -0.88 mm and -1.03 mm (F= -0.04,P=0. 555 ), for grey registration ,x: -0.32 mm, -0.341 mm, -0.395 mm and - 0.37 mm(F=-0.01, P=0.874);y:0.34 mm,0.54 mm, -0.04 mm and 0.27 mm (F= -0.03,P=0.622);x:-1.12 mm,- 1.15 mm, - 1.13 mm and - 1.04 mm (F=0. 00,P=0. 812). Conclusions With the same registrated box and imagine quality, the location of the lesions in lung, registred methods and different doctors are not the influencing factors for CBCT imagine registration.